Combination treatment for multiple sclerosis

ABSTRACT

The present invention provides for the treatment of multiple sclerosis by combination therapies of CD52 specific antibodies, such as CAMPATH-1H, with either a Type 1 interferon or coploymer-1.

FIELD OF THE INVENTION

The present invention relates to the use of CD52 specific antibodies inthe treatment of autoimmune diseases. More particularly it relates tothe use of anti-CD52 antibodies in combination with other therapeuticagents in the treatment of multiple sclerosis.

BACKGROUND OF THE INVENTION

Multiple sclerosis (“MS”) is a chronic and progressive inflammatorydisease of the central nervous system (“CNS”). The disease ischaracterized by loss of the myelin layer that insulates nerve fibersand, while the precise cause of MS is unknown, it is generallyrecognized that there is an autoimmune component in the etiology of thedisease. The areas of demyelination, or plaques, are inflammatory innature with infiltration by T and B-lymphocytes and macrophages. Theremay also be a decrease in T-suppressor cell number and function(Ffrench-Constant, 1994). The degree of inflammation correlates with theamount of axonal loss. Chronic lesions lose their components in natureand are characterized by demyelinated axons separated by a dense networkof astrocyte processes (Trapp et al., 1998).

Patients with multiple sclerosis can be generally categorized intodifferent groups (Weinshenker, 1995). Relapsing remitting multiplesclerosis (“RR-MS”) is characterized by periodic exacerbations of MSsymptoms followed by periods of remission with complete or near-completerecovery. In secondary progressive multiple sclerosis (“SP-MS”) thereare distinct attacks as in RR-MS, but the intervening recovery isincomplete and there is a progression of disability in the “recovery”phases. In primary progressive multiple sclerosis (“PP-MS”), there is arapid deterioration in condition to severe disability with nodiscernable periods of remission.

There is currently no cure for MS, although a number of drugs are usedto treat the symptoms of the disease. Interferon-β is one of the mosteffective drugs available that can inhibit the progression of thedisease (Smith & Darlington, 1999). Glatiramer acetate/copolymer I(Copaxone®) has also shown ability to inhibit disease progression inRR-MS (Johnson et al., 1998). Corticosteroids, particularlymethylprednisolone, have been used extensively in the treatment ofrelapses and acute exacerbations (Bansil et al., 1995). Numerous otherdrugs have been used for specific aspects of symptomatic treatment, suchas for fatigue and motor, urinary, sensory and psychological symptoms(Smith & Darlington, 1999).

While there are a number of treatment modalities for MS, there is acurrent need for improved treatments for this chronic disabling disease.

SUMMARY OF THE INVENTION

The present invention provides for the treatment of MS by combinationtherapies comprising administration of CD52-specific antibodies witheither a Type 1 interferon or copolymer-1, such combination therapiesproviding greater benefits than those associated with the correspondingsingle-agent treatment regimens. One aspect of the invention providesfor a method for the treatment of an MS patient comprising administeringan anti-CD52 antibody as a first treatment modality and Type 1interferon as a second treatment modality, wherein the combinedadministration is effective to treat MS. The Type 1 interferon may be aninterferon-α or interferon-β. In a preferred embodiment, the anti-CD52antibody is CAMPATH-1H. In some embodiments, the interferon-β is aninterferon-β-1b, which may be Rebif®D or Avonex® or the interferon-β isan interferon-β-1b, which may be Betaseron®. The interferon-α may beinterferon-α2a which may be ROFERON®, or the interferon-α may beinterferon-α2b which may be INTRON® A or PEG-INTRON™.

Another aspect of the invention provides for a method for the treatmentof an MS patient comprising administering an anti-CD52 antibody as afirst treatment modality and copolymer-1 as a second treatment modality,wherein the combined administration is effective to treat MS. In apreferred embodiment, the anti-CD52 antibody is CAMPATH-1H. In someembodiments, the copolymer-1 is Copaxone®.

DETAILED DESCRIPTION OF THE INVENTION

A. CD52 Specific Antibodies

The CD52 (CAMPATH-1) antigen is a glycoprotein expressed on lymphocytes,monocytes, macrophages, NK cells, and tissues of the male reproductivesystem (Hale et al., 1990). Antibodies to CD52 are disclosed in U.S.Pat. No. 5,846,534, herein incorporated by reference. The use of CD52specific antibodies for the treatment of MS is disclosed by U.S. Pat.No. 6,120,766, herein incorporated by reference. Anti-CD52 antibodiesbind to all lymphocytes, a majority of monocytes, macrophages, and NKcells, and a subpopulation of granulocytes, but lyse only lymphocytes invivo. CAMPATH-1M is a rat IgM monoclonal antibody that has been usedextensively to deplete T-cells in bone marrow harvests prior totransplantation. CAMPATH-1G is a rat IgG2b class-switch variant of aIgG2a antibody. This antibody has been used in vivo forimmunosuppression in transplant patients. CAMPATH-1H is a humanizedmonoclonal antibody and is approved for the treatment of B-cell chroniclymphocytic leukemia in patients who have been treated with alkylatingagents and who have failed fludarabine therapy. CAMPATH-1H isdistributed as CAMPATH® (Alemtuzumab) in the U.S. (Berlex) andMABCAMPATH™ in Europe (Schering A. G.).

Infusion of CAMPATH-1H results in the rapid fall of lymphocyte andmonocyte counts over the first hour post-treatment and a prolongedlymphopenia that ensues for over 2 years. Administration of CAMPATH-1Hto patients with SP-MS resulted in a sustained and marked decrease inactive inflammation and the prevention of new symptoms, although somepatients experienced progressive disability and increasing brain atrophyassociated with axonal loss (Paolillo et al., 1999; Coles et al., 1999).Axonal degradation correlated with the extent of cerebral inflammationin the pretreatment phase.

B. Interferon-β

There are three recombinant interferon-β products available for thetreatment of MS: Rebif® (Serono); Avonex® (Biogen); and Betaseton®(Bertex). The later is a mutant lacking the N-terminal methionine andserine substituted for cysteine at the 17 position. Betaseron® is madein Escherichia coli and lacks the glycosylation of the native moleculeand is thereby classified as an interferon-β-1b. Rebif® and Avonex® havethe native interferon-β sequence and being produced in CHO cells areglycosylated and are thereby termed as interferon-β-1a's. The term“interferon-(3” as used herein encompasses both interferon-β-1a andinterferon-β-1b variants.

In RR-MS patients, interferon-β decreases both the development oflesions and the occurrence of new lesions (Simon et al., 1998; Stone etal., 1997; Calabresi et al., 1997) and also has been reported to improvecognitive function (Pliskin et al., 1996). In SP-MS, interferon-β hasbeen shown to delay the sustained neurological deteriorationcharacteristic of these patients (European Study Group on Interferonβ-1b in Secondary Progressive MS, 1998).

Although the mode of action of interferon-β remains uncertain, oneproposed mechanism is via inhibition of the immunological effects andsynthesis of interferon-γ (Pantich & Bever, 1993; Corsini et al., 1997;Peitereit et al., 1997). However, type 1 interferons (interferons α andβ) may directly upregulate T-cell interferon-γ production (Karp et al.,2001), a conclusion that is consistent with observations that bothcirculating neopterin levels and MHC-Class II molecule expression oncirculating monocytes rise when patients are started on interferon-β(Chiang et al., 1993; Spears et al., 1987). Further, the number ofcirculating interferon-γ secreting cells are increased in the first twomonths of treatment with interferon-β (Arnason et al., 1997). Analternative mechanism for interferon-β activity is via upregulation ofinterleukin-10 and downregulation of interleukin-12, resulting indecreased presentation of antigens implicated in demyelination and lossof oligodendrocytes associated with MS (Karp et al., 2001). The actionof interferon-β on MS via modulation of theinterleukin-10/interleukin-12 axis may be compromised by the directupregulation of interferon-γ production. Interleukin-10, the dominantendogenous inhibitor of interleukin-12, is produced by a wide variety ofcells, including antigen presenting cells, astrocytes and microglia inaddition to T-cells. Interleukin-12 is produced predominantly by antigenpresenting cells and a subset of B cells and to a lesser extent byastrocytes and microgoia (Karp et al., 2001). Consequently, while thepresent invention is not bound by any one theory, theinterferon-β-induced interferon-γ production may be negated by removalof the interferon-γ T-cells by administration of anti-CD52 antibody,while leaving the interleukin10/interleukin-12 axis functional andthereby potentiating the efficacy of interferon-β.

C. Interferon-α

Interferon-α products currently marketed include ROFERON®, a recombinantinterferon-α2a marketed by Hoffman-La Roche, Nutley N.J., and INTRON® A,a recombinant interferon-α2b marketed by Schering Corp., Kenilworth N.J.Schering also markets PEG-INTRON™, an interferon-α2b conjugated withmonomethoxy polyethylene glycol (“PEG”), called.

In RR-MS patients, interferon-α2a reduces exacerbation rate and MRIsigns of disease activity (Durelli et al., 1994; Durelli et al., 1996;Myhr et al., 1999). The immunomodulatory effects of interferon-α arebroadly similar to those described above for interferon-β(Weinstock-Guttman et al., 1995; Panitch & Bever, 1993; Durelli et al.,1994; Bongioanni et al., 1996; Piazzolla et al., 2000; Byrnes et al.,2001)

D. Copolymer-1

Copolymer-1 (glatiramer acetate), a synthetic peptide analogue of myelinbasic protein (BP), is a standardized mixture of L-glutamic acid,L-lysine, L-alanine and L-tyrosine with a molar ratio of0.14:0.34:0.43:0.1 and a molecular mass of 4.7-11.0 KDa Copolymer-1 hasbeen demonstrated to have beneficial effects on MRI-defined brainlesions and reduces the relapse rate and accumulated disability of RR-MSpatients (Johnson et al., 2000, Johnson et al., 1998; Mancardi et al.,1998). This agent has been approved for the treatment of RR-MS in theU.S. and is distributed under the name Copaxone® (Teva Pharmaceuticals).The composition and preparation of copolymer-1 are disclosed in U.S.Pat. Nos. 5,981,589 and 6,054,430, both herein incorporated byreference.

Copolymer-1 is cross-reactive with MBP and may inhibit the immuneresponse in MS to 4 MBP, proteolipid protein and/or myelinoilgodendrocyte glycoprotein (Arnon et al., 1996; Ben-Nun et al., 1996;Lea & Goa, 1996). A shift from a Th1-biased T-cell cytokine profile(II-2, IFN-γ, TNF-α) towards a Th2-biased cytokine profile (IL-4, IL-5,IL-6, IL-10, TFG-β) has been observed in copolymer-1 treated MS patients(Duda et al., 2000; Neuhaus et al., 2000). The copolymer-1 specificTh2-type T-cells may be involved in the copolymer-1-induced therapeuticeffects in MS (Aharoni et al., 2000).

D. Formulations and Administration

The pharmaceutical compositions according to the present invention areprepared conventionally, comprising substances that are customarily usedin pharmaceuticals, e.g., Remington's Pharmaceutical Sciences, 18th ed.,Mack Publishing Company (1990), including excipients, carriers,adjuvants, and buffers. The compositions can be administered, e.g.,parenterally, enterally, orally, intramuscularly, subcutaneously,intravenously, by aerosol, intrathecally directly into the cerebralspinal fluid of the CNS, or other routes useful to achieve an effect.For example: anti-CD52 antibodies, preferably CAMPATH-1H, can be givenintravenously (Cloes et al., 1999; Moreau et al., 1996; Moreau et al.,1994, all herein incorporated by reference) and subcutaneously(Schnitzer et al., 1997; Bowen et al., 1997, both herein incorporated byreference); interferon-β may be given subcutaneously (Stone et al.,1997, herein incorporated by reference) and by intramuscularadministration (Simon et al., 1998, herein incorporated by reference);interferon-α may be given subcutaneously (Durelli et al., 1994, hereinincorporated by reference) and by intramuscular administration (Myhr etal., 1999, herein incorporated by reference); and copolymer-1 can beadministered subcutaneously (Johnson et al., 2000, herein incorporatedby reference) and intramuscularly (Jacobs et al., 1994, hereinincorporated by reference).

Conventional excipients include pharmaceutically acceptable organic orinorganic carrier substances suitable for parenteral, enteral, ortopical application that do not deleteriously react with the agents.Suitable pharmaceutically acceptable adjuvants include, but are notlimited to water, salt solutions, alcohols, gum arabic, vegetable oils,polyethylene glycols, gelatine, lactose, amylose, magnesium stearate,talc, silicic acid, viscous paraffin, perfume oil, fatty acidmonoglycerides and diglycerides, pentaerythritol fatty acid esters,hydroxy-methylcellulose, polyvinyl pyrrolidone, cyclodextrins, etc. Thepharmaceutical preparations can be sterilized and, if desired, mixedwith stabilizers, wetting agents, emulsifiers, salts for influencingosmotic pressure, buffers, coloring, flavoring and/or aromaticsubstances, etc., that do not react deleteriously with the activecompounds.

For parenteral application, particularly suitable are injectable sterilesolutions, preferably oil or aqueous solutions, as well as suspensions,emulsions or implants, including suppositories. Ampules are convenientunit dosages.

The compositions can also be formulated in an aqueous solution,optionally with the addition of additives customary in galenicals, forexample, buffers; electrolytes such as sodium chloride; antioxidantssuch as ascorbic acid; adjuvants, e.g., methylcellulose, lactose andmannitol and/or surfactants, e.g., lecithins and Tweens and/or aromaticsubstances for flavoring, e.g., ethereal oils.

Dosage levels and treatment regimens for MS of interferon-β, includingRebif®, Avonex®, and Betaseron®, interferon-α, including ROFERON®,INTRON® A and PEG-INTRON™, and copolymer-1 preparations, includingCopaxone®, are known in the art. E.g., the package insert instructionsfor Rebif indicate a subcutaneous dose of 44 mcg 3 times per week. Aclinical study may suitably use an initial titration e.g., in the first6 weeks Rebif is administered subcutaneously at 11 mcg 3 times per weekfor the first two weeks, 22 mcg 3 times a week for the next two weeks,and 33 mcg for the following two weeks. A patient that has an adversereaction to Rebif may suitably have dose adjustments made in accordanceto the physician's discretion in accordance with the clinical guidelinesprovided by the manufacturer. ROFERON may suitably be given in the rangeof about 4.5 mIU to about 9 mIU subcutaneoulsy or intramuscularly threetimes per week. Copoaxone may suitably be administered subcutaneously ata daily dose of 20 mg.

In some embodiments, the dosage of Type 1 interferons, i.e.,interferon-α or interferon-β, when used in a combination regiment withan anti-CD52 antibody is reduced compared to the interferon-β dosageused in a single-agent treatment regimen. The dosage of a course ofanti-CD52 antibodies, preferably CAMPATH-1H, may vary with the status ofthe MS patient and will generally be in the range of about 10 to about150 mg for an adult patient, usually administered over a period from 1to about 20 days. The course of treatment may be given once or may berepeated at about 3 month, or about six month, or at about 9 month, orabout 12 month, or about 18 month or at about 24 month intervals, thenumber of courses of treatment depending upon the medical status of thepatient, including but not limited, to the symptoms of MS and extent andpersistence of lymphopenia. In some embodiments of the presentinvention, the dosage schedules suitably utilized in a clinical studyare a low dose level of a total of 0.37 mg/kg, a mid dose level of atotal of 0.75 mg/kg and a high dose level of a total of 1.50 mg/kg, allgiven IV over a total of 5 consecutive, i.e., 0.07, 0.15 and 0.30mg/kg/day respectively. Re-treatment is given at months 24 and 48 monthsat a low dose level of a total of 0.22 mg/kg, a mid 4 dose level of atotal of 0.45 mg/kg and a high dose level of a total of 0.90 mg/kg, allgiven IV over a total of 3 consecutive, i.e., 0.07, 0.15 and 0.30mg/kg/day respectively.

The first course of CAMPATH-1H treatment has been associated with areversible exacerbation of existing neurological symptoms and activationof asymptomatic lesions caused by an antibody-induced release ofcytokines (Moreau et al., 1996; Wing et al., 1996). Thiscytokine-release syndrome can be prevented by pretreatment withmethylprednisolone (Coles et al., 1999, herein incorporated byreference).

In accordance with the methods of the present invention, the twotreatment modalities in the combination of anti-CD52 antibodies withinterferon-α, interferon-β or copolymer-1 can be administered separatelyat different times during the course of therapy or concurrently individed or single combination forms. In a preferred embodiment, theadministration of an anti-CD52 antibody precedes the administration ofinterferon-β interferon-α. According to the instant invention, the termadministering is to be understood as embracing all such regimes ofsimultaneous or alternating treatment and the scope of combinations ofanti-CD52 antibodies with interferon-α, interferon-β or copolymer-1includes, in principle, any combination useful for treating MS.

EXAMPLES OF THE INVENTION

A. Clinical Evaluation of Combined Therapies

Patients will be evaluated for clinically definite MS (Poser et al.,1983, herein incorporated by reference). SP-MS patients will have aKurtzke expanded disability status score (EDSS; Kurtzke, 1983, hereinincorporated by reference) of between 3.0-7.0 inclusive, with a recordedhistory of a 1.0 point or more increase in the previous 2 years.Immunosuppressive or immunomodulatory treatment or other putativetreatments for MS are not permitted for a defined period prior to entryinto the trial. Suitable eligibility criteria are provided by Polman etal., 1995, herein incorporated by reference. RR-MS patients will havehad at least two relapses within the last two years and to be free fromsteroid use for at least two months. Suitable treatment cohorts forCAMPATH-1H and interferon-β combined therapy include: (1) treatment withCAMPATH-1H; (2) treatment with interferon-β; and (3) treatment withCAMPATH-1H and interferon-β. Suitable treatment cohorts for CAMPATH-1Hand interferon-α combined therapy include: (1) treatment withCAMPATH-1H; (2) treatment with interferon-α; and (3) treatment withCAMPATH-1H and interferon-α. Suitable treatment cohorts for CAMPATH-1Hand copolymer-1 combined therapy include: (1) treatment with CAMPATH-1H;(2) treatment with copolymer-1; and (3) treatment with CAMPATH-1H andcopolymer-1.

Efficacy of treatments are suitably monitored by MRI, wherein MRIstudies are calculated at baseline and followed up periodically over aperiod of up to 5 years. MRI can obtain images that are proton density(PD) weighted, T1-weighted, and T2 weighted (Paty, 1993; Francis et al.,1995, both herein incorporated by reference). Suitable MRI measurementtechniques/parameters include: brain volume; gadolinium enhancement forthe evaluation of disruption of the blood-brain barrier andinflammation; evaluation of new T2 lesions as an indicator ofinflammation; enlarging T2 lesions as an indicator of increasinginflammation; and T1 “black holes” for permanent demyelination andaxonal loss (Adams et al., 1999; Coles et al., 1999; Simon et al., 1998;Bruck et al., 1997; Katz et al., 1993; Hawkins et al., 1991, all hereinincorporated by reference). MS-related disability is evaluated over thesame period according to the Kurtzke EDSS system. One efficacy endpointis the proportion of patients without sustained accumulation ofdisability (“SAD”) at defined time points after initiation of treatment.SAD is defined as an increase of ≧1.0 point of the EDSS sustained over asix-month consecutive period. Other criteria of efficacy include numberof relapses, time to first relapse and rate of cerebral atrophy.

B. Preclinical Evaluation of Combined Therapies

1. Mouse Model

Preclinical evaluation is performed in an experimental autoimmuneencephalomyelitis (“EAE”) model, suitable antigens being myelinoligodendrocyte glycoprotein (“MOG”), myelin basic protein (“MBP”) andproteolipid protein (“PLP”). Female mice (e.g., 6 to 16 weeks of age) ofa strain susceptible to EAE induction are used e.g., SLJ/J (u et al.,2001), (SWR×SJL/J)F₁ (Yu et al., 1996, Sobel et al., 1991),(SJL/J×BALB/c)F₁ (Aharoni et al., 2000), C57BL/6 (Zhang et al., 2002) orPL/J (Soos et al., 2002). Immunization is suitably carried out asdetailed in the art (e.g., Du et al., 2001; Yu et al., 1996; Sobel etal., 1991; Aharoni et al., 2000; Zhang et al., 2002; Soos et al., 2002and Gold et al., 2000, all herein incorporated by reference). Effectivedoses of interferon-α, interferon-β and copolymer-1 have been describedin mouse models (Brod et al., 1995; Yu et al., 1996; Aharoni et al.,2000, all herein incorporated by reference).

Anti-mouse CD52 is given subcutaneously for a number of consecutive days(e.g., 5 days at days 8-13). Effective dose range is established bymonitoring the depletion of CD52 positive cells, e.g., T-cells. The B7antigen (B7-Ag) is the mouse homolog to CD52 (Tone et al., 1999), and arat anti-mouse B7-Ag IgG2a monoclonal antibody is described by Kubota etal. (1990). Preferably, an IgG2b class-switch variant is isolated, e.g.,by sib selection using red cell-linked antibodies to identify thedesired secreted Ig by reverse passive haemagglutination (Hale et al.,1987, herein incorporated by reference).

Cohorts include control (vehicle only) and single agents and combinedtreatments of anti-mouse CD52 and either interferon-β or copolymer 1.

Mice are graded for clinical signs of EAE according to a suitableguideline, e.g., Grade 1—tail weakness or tail paralysis; Grade 2—hindleg paraparesis or hemiparesis; Grade 3—hind leg paralysis orhemiparalysis; Grade 4—complete paralysis (tetraplegy), moribund state,or death. Ataxia will be routinely assessed. A disease remission isdefined as an improvement in disease score from either 3 or 4 to 1, orfrom 2, 3 or 4 to 0, that is maintained for at least 2 consecutive days.A relapse is defined as an increase in the clinical deficit of at leasttwo points that lasted for at least 2 days. Body weight is measuredpretest and daily through the test.

Clinical pathology comprises lymphocyte proliferative responses aresuitably assessed in viable spleen cells, inguinal lymph node cells orperipheral blood mononuclear cells in mice sacrificed at suitable timepoints, e.g., days 15 and 60.

Histopathology comprises evaluation of brain and spinal cord from allrats. Inflammatory index is determined from the number of perivascularinflammatory infiltrates of each animal on an average of 15 completecross sections of spinal cord. The degree of myelination is suitablyevaluated separately for brain and spinal cord and scored, e.g., 0.5traces of perivascular or subpial demyelination; 1=marked perivascularor subpvial demyelination; 2=confluent perivascular or subpialdemyelination; 3=massive confluent demyelination (e.g., half of spinalcord, one complete optic nerve); and 4=extensive demyelination(transverse myelitis, half of the cerebellar white matter or more, bothcomplete optic nerves).

2. Rat Model

Preclinical evaluation is again performed in an experimental autoimmuneencephalomyelitis (“EAE”) model, suitable antigens being myelinoligodendrocyte glycoprotein (“MOG”), myelin basic protein (“MBP”) andproteolipid protein (“PLP”). Female rats (10 to 14 weeks of age) of astrain susceptible to EAE induction are used (e.g., DA, Lewis.1A,Lewis.AV1 or Lewis.1N-fulminant disease model). Rats are immunizedutilizing a suitable regimen, e.g., intradermal injection at the base ofthe tail, e.g., 1, 5, 20, 50 or 100 μL, of an MOG inoculum, e.g., 1:1MOG and saline emulsified with CFA (Sigma Chemical Co, ST Louis Mo.)containing 200 μg of Mycobacterium tuberculosis (strain H 37 RA, DifcoLabs Irvine Calif.).

Anti-rat antibodies are raised against the rat B7-antigen homolog toCD52 (Kirchhoff, 1994, Eccleston et al., 1994), by methods generallyknow in the art. Anti-rat CD52 antibody is given subcutaneously for anumber of consecutive days (e.g., 5 days at days 8-13 post-inoculation).Interferon-β is given by subcutaneous dose at a suitable starting point,e.g., day 8, and then every other day thereafter. Copolymer-1 is givenby subcutaneous dose on a suitable starting point, e.g., day 8, and thendaily thereafter. Cohorts include control (vehicle only) and singleagents and combined treatments of anti-rat CD52 and eitherinterferon-βor copolymer-1.

Rats are graded for clinical signs of EAE according to a suitableguideline, e.g.: Grade 1—tail weakness or tail paralysis; Grade 2—hindleg paraparesis or hemiparesis; Grade 3—hind leg paralysis orhemiparalysis; Grade 4—complete paralysis (tetraplegy), moribund state,or death. Ataxia will be routinely assessed. A disease remission isdefined as an improvement in disease score from either 3 or 4 to 1, orfrom 2, 3 or 4 to 0, that is maintained for at least 2 consecutive days.A relapse is defined as an increase in the clinical deficit of at leasttwo points that lasted for at least 2 days. Body weight is measuredpretest and daily through the test.

Clinical pathology comprises lymphocyte proliferative responses aresuitably assessed in viable spleen cells, inguinal lymph node cells orperipheral blood mononuclear cells in rats sacrificed at suitabletimepoints, e.g., days 15 and 60.

Histopathology suitably comprises evaluation of brain and spinal cordfrom all rats. Inflammatory index is determined from the number ofperivascular inflammatory infiltrates of each animal on an average of 15complete cross sections of spinal cord. The degree of myelination willbe evaluated separately for brain and spinal cord and scored, e.g.:0.5=traces of perivascular or subpial demyelination; 1=markedperivascular or subpvial demyelination; 2=confluent perivascular orsubpial demyelination; 3=massive confluent demyelination (e.g., half ofspinal cord, one complete optic nerve); and 4 extensive demyelination(transverse myelitis, half of the cerebellar white matter or more, bothcomplete optic nerves).

The present invention has been shown by both description and examples.The examples are only for exemplification and cannot be construed tolimit the scope of the invention. One of ordinary skill in the art willenvision equivalents to the inventive process described by the followingclaims that are within the scope and spirit of the claimed invention.

REFERENCES

The following references, to the extent that they provide exemplaryprocedural or other details supplementary to those set forth herein, arespecifically incorporated herein by reference.

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1. A method for the treatment of a human subject suffering from multiplesclerosis, comprising administering to said subject an anti-CD52antibody as a first treatment modality and a Type 1 interferon as asecond treatment modality, wherein the administration of said anti-CD52Yb antibody when combined with the administration of said Type 1interferon-β is effective to treat said multiple sclerosis.
 2. Themethod of claim 1, wherein said anti-CD52 antibody is CAMPATH-1H.
 3. Themethod of claim 1, wherein said Type 1 interferon is interferon-β. 4.The method of claim 3, wherein said interferon-β is interferon-β-1a orinterferon-β-1b.
 5. The method of claim 4, wherein said interferon-β-1ais Rebif® or Avonex®.
 6. The method of claim 4, wherein saidinterferon-β-1b is Betaseron®.
 7. The method of claim 3, wherein saidType 1 interferon is interferon-α.
 8. The method of claim 7, whereinsaid interferon-α is interferon-α2a or interferon-α2b.
 9. The method ofclaim 8, wherein said interferon-α2a is ROFERON®.
 10. The method ofclaim 8, wherein said interferon-α2b is INTRON® A or PEG-INTRON™.
 11. Amethod for the treatment of a human subject suffering from multiplesclerosis, comprising administering to said subject an anti-CD52antibody as a first treatment modality and copolymer-1 as a secondtreatment modality, wherein the administration of said anti-CD52antibody when combined with the administration of said copolymer-1 iseffective to treat said multiple sclerosis.
 12. The method of claim 11,wherein said anti-CD52 antibody is CAMPATH-1H.
 13. The method of claim12, wherein said copolymer-1 is Copaxone®.